Exosomal miR-221/222 enhances tamoxifen resistance in recipient ER-positive breast cancer cells
Exosomal miR-221/222 enhances tamoxifen resistance in recipient ER-positive breast cancer cells
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外泌体 miR-221/222 增强受体 ER 阳性乳腺癌细胞对他莫昔芬的耐药性。
DOI：
10.1007/s10549-014-3037-0
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发表时间：
2014-09-01
影响因子：
3.8
通讯作者：
Zhang, Jian
中科院分区：
文献类型：
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作者：
Wei, Yifang;Lai, Xiaofeng;Zhang, Jian
最近的研究表明，特定的 miRNA，例如 miR-221/222，可能是乳腺癌中他莫昔芬耐药的原因。封装在外泌体中的分泌 miRNA 可以充当细胞间生物信使。我们的目的是研究分泌的 miR-221/222 在 ER 阳性乳腺癌细胞他莫昔芬耐药中的作用。进行透射电子显微镜分析和纳米颗粒跟踪分析以确定 MCF-7(TamR)（他莫昔芬耐药）和 MCF-7(wt)（他莫昔芬敏感）细胞之间的外泌体差异。采用PKH67荧光标记法检测MCF-7(TamR)细胞来源的外泌体进入MCF-7(wt)细胞。通过细胞活力、细胞凋亡和集落形成来分析外泌体对他莫昔芬耐药性传播的潜在功能。使用miRNA微阵列和qPCR来检测和比较两种细胞和外泌体中的miRNA表达水平。作为 miR-221/222 的靶标，p27 和 ER α 通过蛋白质印迹和 qPCR 进行分析。与MCF-7(wt)外泌体相比，MCF-7(TamR)外泌体的浓度和大小分布存在显着差异。与 MCF-7(TamR) 细胞相比，MCF-7(wt) 细胞的外泌体 RNA 和蛋白质含量增加。 MCF-7(TamR)外泌体可以进入MCF-7(wt)细胞，然后释放miR-221/222。 miR-221/222升高有效降低了P27和ERα靶基因的表达，从而增强了受体细胞对他莫昔芬的耐药性。我们的结果首次表明，分泌的 miR-221/222 可作为信号分子介导他莫昔芬耐药性的通讯。
Recent studies have demonstrated that specific miRNAs, such as miR-221/222, may be responsible for tamoxifen resistance in breast cancer. Secreted miRNAs enclosed in exosomes can act as intercellular bio-messengers. Our objective is to investigate the role of secreted miR-221/222 in tamoxifen resistance of ER-positive breast cancer cells. Transmission electron microscopy analysis and nanoparticle tracking analysis were performed to determine the exosomes difference between MCF-7(TamR) (tamoxifen resistant) and MCF-7(wt) (tamoxifen sensitive) cells. PKH67 fluorescent labeling assay was used to detect exosomes derived from MCF-7(TamR) cells entering into MCF-7(wt) cells. The potential function of exosomes on tamoxifen resistance transmission was analyzed with cell viability, apoptosis ,and colony formation. MiRNA microarrays and qPCR were used to detect and compare the miRNAs expression levels in the two cells and exosomes. As the targets of miR-221/222, p27 and ER alpha were analyzed with western blot and qPCR. Compared with the MCF-7(wt) exosomes, there were significant differences in the concentration and size distribution of MCF-7(TamR) exosomes. MCF-7(wt) cells had an increased amount of exosomal RNA and proteins compared with MCF-7(TamR) cells. MCF-7(TamR) exosomes could enter into MCF-7(wt) cells, and then released miR-221/222. And the elevated miR-221/222 effectively reduced the target genes expression of P27 and ER alpha, which enhanced tamoxifen resistance in recipient cells. Our results are the first to show that secreted miR-221/222 serves as signaling molecules to mediate communication of tamoxifen resistance.